Development of Gene Therapy & Gene Editing for Lung Disorders

Inherited diseases of the lung such as Surfactant Protein deficiencies are disorders that typically present shortly after birth, resulting in alveolar collapse and respiratory distress in neonates. The prognosis can be poor and in many cases the disorder is fatal, with affected babies dying shortly after delivery. Furthermore, due to the lack of donor organs, and the unstable state of the disease, lung transplantation is rarely attempted in such young children. Progress in developing treatments has been slow as the majority of such diseases are very rare, with many causative mutations and disparate mechanisms of action.

We propose to apply gene therapy and gene editing technologies to repair the mutant genes in lung cells shortly after birth. The full correction of such disorders will be technically very challenging, but we hypothesise that even partial correction may have a significant effect on patient outcome, for example by extending the window in which lung transplantation could be offered. We have experience of the design, production and use of two gene delivery technologies for the treatment of lung diseases: viral vectors based on Lentivirus (LV) and Adeno-Associated Virus (AAV). In order to treat cystic fibrosis, a more common lung disorder, we have developed a highly potent, third-generation, self-inactivating simian immunodeficiency viral vector. The natural viral envelope protein has been replaced with the F & HN proteins from Sendai virus – a practice known as pseudotyping – to direct highly efficient gene delivery to a range of lung cell types. Recombinant AAV is available in a large number of serotypes that offer highly specific cell targeting. This project will exploit these lung gene delivery vectors in combination with cell-specific promoters, to direct long-lasting transgene expression in target cells. The vectors will be evaluated for the potential to reverse disease features in primary human lung cell cultures and mouse models for the treatment of lung disorders.

TRAINING OPPORTUNITIES

Training Opportunities: The project will be based in the Gene Medicine Research Group in the John Radcliffe Hospital. The group is expert in the development of gene therapy for lung diseases, and has experience in conducting clinical gene therapy trials and vector manufacturing, exposing students to all aspects of translational research. The student will receive training in techniques such as: molecular biology, gene editing, cell culture, microscopy & imaging, protein characterisation along with virus production/purification and functional evaluation, PCR, FACS, Western blotting, immunocytochemistry, ELISA, Quantitative (RT)-PCR, lentivirus production, & Tangential Flow Filtration (TFF) methods.

As well as the specific training detailed above, students will have access to a wide-range of seminars and training opportunities through the many research institutes and centres based in Oxford. Students are also able to attend the Methods and Techniques course run by the MRC Weatherall Institute of Molecular Medicine. This course runs through the year, ensuring that students have the opportunity to build a broad-based understanding of differing research techniques.

Generic skills training is offered through the Medical Sciences Division’s Skills Training Programme. This programme offers a comprehensive range of courses covering many important areas of researcher development: knowledge and intellectual abilities, personal effectiveness, research governance and organisation, and engagement, influence and impact. Students are actively encouraged to take advantage of the training opportunities available to them.

The Department has a successful mentoring scheme, open to graduate students, which provides an additional possible channel for personal and professional development outside the regular supervisory framework. We hold an Athena SWAN Silver Award in recognition of our efforts to support the careers of female students and staff.